What is the difference between microbial identification and profiling? Identification is determining the microbe’s presence or absence in your sample which requires running a No Template Control during your analysis. Profiling is determining the microbe’s relative expression in two or more experimental conditions for which you will need to run a reference sample and a normalizer (provided by QIAGEN).

Specificity

Each Microbial DNA qPCR Assay is stringently tested to ensure that it detects only one target species or gene (see figure, Microbial DNA qPCR Assays are highly specific). For assays that do detect more than one target, a list of detected targets and in silico predictions is included on the product sheet.

Microbial DNA qPCR Assays are designed to detect bacterial 16S rRNA gene and fungal ribosomal rRNA gene sequences for species identification, as well as detecting virulence factor genes and antibiotic resistance genes using PCR amplification primers and hydrolysis-probe detection.

Procedure

The procedure for Microbial DNA qPCR Multi-Assay Kits is simple, and can be performed in any laboratory with a real-time PCR instrument. DNA is isolated from the sample using the appropriate QIAamp kit, and PCR reactions are set up using the appropriate Microbial qPCR Mastermix for the PCR instrument. Three control reactions are prepared for each sample, including Positive PCR Control, No Template Control, and the Microbial DNA Positive Control, as well as the Microbial DNA qPCR Assays. Real-time PCR is performed and data is analyzed using web-based data analysis software or Excel templates.

Applications

Microbial DNA qPCR Multi-Assay Kits are highly suited for the detection of related bacterial or fungal species, or microbial genes for antibiotic resistance or virulence factors, from a variety of samples.